Characterisation of new intracellular membranes in Escherichia coli accompanying large scale over-production of the b subunit of F₁F_o ATP synthase

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• 作者：Arechaga ; Ignacio ; Miroux ; Bruno ; et. al.
• 关键词：Over-expression ; Membrane protein ; Intracytoplasmic membrane ; Escherichia coli ; IPTG ; isopropyl-2-D-thio-galactopyranoside ; TLC ; thin layer chromatography ; HPLC ; high performance liquid chromatography ; e.s.i.-m.s. ; electrospray ionisation-mass spectrometr
• 刊名：FEBS Letters
• 出版年：2000
• 出版时间：October 6, 2000
• 年：2000
• 卷：482
• 期：3
• 页码：215-219
• 全文大小：925 K

文摘

Recombinant membrane proteins in Escherichia coli are either expressed at relatively low level in the cytoplasmic membrane or they accumulate as inclusion bodies. Here, we report that the abundant over-production of subunit b of E. coli F₁F_o ATP synthase in the mutant host strains E. coli C41(DE3) and C43(DE3) is accompanied by the proliferation of intracellular membranes without formation of inclusion bodies. Maximal levels of proliferation of intracellular membranes were observed in C43(DE3) cells over-producing subunit b. The new proliferated membranes contained all the over-expressed protein and could be recovered by a single centrifugation step. Recombinant subunit b represented up to 80 % of the protein content of the membranes. The lipid:protein ratios and phospholipid compositions of the intracellular membranes differ from those of bacterial cytoplasmic membranes, and they are particularly rich in cardiolipin.